{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,15]],"date-time":"2026-04-15T05:55:50Z","timestamp":1776232550951,"version":"3.50.1"},"reference-count":26,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,12,10]],"date-time":"2024-12-10T00:00:00Z","timestamp":1733788800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003725","name":"National Research Foundation of Korea","doi-asserted-by":"publisher","award":["2023R1A2C1007319, RS-2023-00258956, 2021R1F1A1060347"],"award-info":[{"award-number":["2023R1A2C1007319, RS-2023-00258956, 2021R1F1A1060347"]}],"id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["P01AI145815"],"award-info":[{"award-number":["P01AI145815"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:p>The functional specialization of CD4 T lymphocytes into various subtypes, including T<jats:sub>H<\/jats:sub>1 and T<jats:sub>FH<\/jats:sub> cells, is crucial for effective immune responses. T<jats:sub>FH<\/jats:sub> cells facilitate B cell differentiation within germinal centers, while T<jats:sub>H<\/jats:sub>1 cells are vital for cell-mediated immunity against intracellular pathogens. Integrin \u03b14, a cell surface adhesion molecule, plays significant roles in cell migration and co-stimulatory signaling. In this study, we investigated the role of Integrin \u03b14 in regulating T<jats:sub>FH<\/jats:sub> and T<jats:sub>H<\/jats:sub>1 cell populations during acute viral infection using CRISPR-Cas9 gene editing. To effectively delete the <jats:italic>Itga4<\/jats:italic> in primary mouse CD4 T cells, we selected various combinations of crRNAs and generated ribonucleoprotein complexes with fluorochrome-conjugated tracrRNAs and Cas9 proteins. These crRNA pairs enhanced gene deletion by generating deletions in the gene. By analyzing the effects of <jats:italic>Itga4<\/jats:italic> deficiency on T<jats:sub>FH<\/jats:sub> and T<jats:sub>H<\/jats:sub>1 cell differentiation during acute LCMV infection, we found that optimized crRNA pairs significantly increased the T<jats:sub>H<\/jats:sub>1 cell population. Our results highlight the importance of selecting and combining appropriate crRNAs for effective CRISPR-Cas9 gene editing in primary CD4 T cells. Additionally, our study demonstrates the role of Integrin \u03b14 in regulating the differentiation of CD4 T cells, suggesting the potential molecular mechanisms driving T cell subset differentiation through integrin targeting.<\/jats:p>","DOI":"10.3389\/fimmu.2024.1445341","type":"journal-article","created":{"date-parts":[[2024,12,10]],"date-time":"2024-12-10T05:11:16Z","timestamp":1733807476000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":2,"title":["Efficient gene deletion of Integrin alpha 4 in primary mouse CD4 T cells using CRISPR RNA pair-mediated 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